Juan L. García

Novel Genomic Imbalances in B-Cell Splenic Marginal Zone Lymphomas Revealed by Comparative Genomic Hybridization and Cytogenetics

Splenic marginal zone lymphoma (SMZL) has recently been recognized in the World Health Organization classification of hematological diseases as distinct type of non-Hodgkins lymphoma. In contrast to the well-established chromosomal changes associated with other B-cell non-Hodgkins lymphoma, few genetic alterations have been found associated with SMZL. The aim of our study was to analyze by comparative genomic hybridization (CGH) the chromosomal imbalances in 29 patients with SMZL and to correlate these findings with clinical and biological characteristics and patient outcome. In 21 cases, cytogenetic studies were simultaneously performed. Most of the patients (83%) displayed genomic imbalances. A total of 111 DNA copy number changes were detected with a median of four abnormalities per case (range, 1 to 12). Gains (n = 92) were more frequent than losses (n = 16), while three high-level amplifications (3q26-q29, 5p11-p15, and 17q22-q25) were observed. The most frequent gains involved 3q (31%), 5q (28%), 12q and 20q (24% each), 9q (21%), and 4q (17%). Losses were observed in 7q (14%) and 17p (10%). SMZL patients with genetic losses had a shorter survival than the remaining SMZL patients (P < 0.05). In summary, chromosomal imbalances in regions 3q, 4q, 5q, 7q, 9q, 12q, and 20q have been detected by CGH in SMZL. Patients with SMZL displaying genetic losses by CGH had a short survival.

Inflammatory myofibroblastic tumor of bone: report of two cases with evidence of clonal chromosomal changes.

Inflammatory myofibroblastic tumor (inflammatory pseudotumor) is a pseudosarcomatous lesion that is recognized with increasing frequency in various anatomic locations. However, this lesion has not been previously reported in bone. We report on two cases of inflammatory myofibroblastic tumor occurring in bone in young adults. Both tumors presented as slightly painful, osteolytic, and well-delineated lesions of the distal femur, with a hyperintense signal on T2-weighted magnetic resonance imaging. The patients had an uneventful recovery after curettage. The follow-up time was 11 months for both patients, and no recurrence was noted. On histologic examination, the lesions were characterized by collagen-rich and generally poorly cellular tissue containing spindled to plump (myo)fibroblast-like cells and a variable admixture of inflammatory cells. Focal calcifications and reactive bone formation were present. Clonal, albeit different, chromosomal changes were found in both cases (47,XY,-9,-12,add(21)(q21),+der(?)t(?;9)(?;q11), +mar,+r and 47, XY, +r/47, idem, add(12)(p13)). The present and other reported cytogenetic findings suggest that inflammatory myofibroblastic tumors could well be neoplastic.

Poplar growth and yield in short rotation coppice: model simulations using the process model SECRETS

Abstract The process model SECRETS was adapted to simulate coppice growth of poplar. The effects of soil type, irrigation, nitrogen fertilization and rotation cycle on growth and yield were studied. Simulated average production on an agricultural soil was 12.4 t ha −1 year −1 . Poplar growth was strongly reduced on sandy soils ( 6 t ha −1 year −1 ). Irrigation increased yield on all soil types by 25% while fertilization increased yield by 26%. The highest yield was simulated for the combined irrigation/fertilization ( 22.45 t ha −1 year −1 ). Length of rotation cycle greatly influenced yield; optimal rotation cycle in terms of yield was 3 or 4 years. The results clearly indicate a useful role for process models in predicting effects of management strategies on poplar yield in short rotation coppice.

Energy budget and greenhouse gas balance evaluation of sustainable coppice systems for electricity production

The use of bio-energy crops for electricity production is considered an effective means to mitigate the greenhouse effect, mainly due to its ability to substitute fossil fuels. A whole range of crops qualify for bio-energy production and a rational choice is not readily made. This paper evaluates the energy and greenhouse gas balance of a mixed indigenous hardwood coppice as an extensive, low-input bio-energy crop. The impact on fossil energy use and greenhouse gas emission is calculated and discussed by comparing its life cycle (cultivation, processing and conversion into energy) with two conventional bio-energy crops (short rotation systems of willow and Miscanthus). For each life cycle process, the flows of fossil energy and greenhouse gas that are created for the production of one functional unit are calculated. The results show that low-input bio-energy crops use comparatively less fossil fuel and avoid more greenhouse gas emission per unit of produced energy than conventional bio-energy crops during the first . Where the mixed coppice system avoids up till eq./GJ, Miscanthus does not exceed eq./GJ. After their performances become comparable, amounting to eq./ha/GJ. However, if the land surface itself is chosen as a functional unit, conventional crops perform better with respect to mitigating the greenhouse effect. Miscanthus avoids a maximum of eq./ha/yr, while mixed coppice attains eq./ha/yr at the most.

A high number of losses in 13q14 chromosome band is associated with a worse outcome and biological differences in patients with B-cell chronic lymphoid leukemia

In B-cell chronic lymphoid leukemia, patients with 13q14 deletion generally have a favorable outcome. The findings of this study suggest that the number of malignant cells with 13q14 deletion may influence the outcome of patients with this cytogenetic abnormality as a single chromosomal aberration. A high number of malignant cells carrying the 13q14 deletion, as assessed by FISH, appears to be associated with short overall survival and time to progression. Background Among patients with B-cell chronic lymphoid leukemia, those with 13q14 deletion have a favorable outcome. However, whether the percentage of cells with 13q- influences the prognosis or the biological characteristics of this disease is unknown. We analyzed the clinico-biological characteristics and outcome of patients with B-cell chronic lymphoid leukemia with loss of 13q as the sole cytogenetic aberration. Design and Methods Three hundred and fifty patients with B-cell chronic lymphoid leukemia were studied. Clinical data were collected and fluorescence in situ hybridization and molecular studies were carried out. In addition, a gene expression profile was obtained by microarray-based analysis. Results In 109 out of the 350 cases (31.1%) loss of 13q was the sole cytogenetic aberration at diagnosis. In the subgroup of patients with 80% or more of cells with loss of 13q (18 cases), the overall survival was 56 months compared with not reached in the 91 cases in whom less than 80% of cells had loss of 13q (p< 0.0001). The variables included in the multivariate analysis for overall survival were the percentage of losses of 13q14 (p=0.001) and B symptoms (p=0.007). The time to first therapy in the group with 80% or more vs. less than 80% of losses was 38 months vs. 87 months, respectively (p=0.05). In the multivariate analysis the variables selected were unmutated status of IgVH (p=0.001) and a high level of β2microglobulin (p=0.003). Interestingly, these differences regarding overall survival and time to first therapy were also present when other cut-offs were considered. The gene expression profile of patients with a high number of losses in 13q14 showed a high proliferation rate, downregulation of apoptosis-related genes, and dysregulation of genes related to mitochondrial functions. Conclusions Patients with B-cell chronic lymphoid leukemia with a high number of losses in 13q14 as the sole cytogenetic aberration at diagnosis display different clinical and biological features: short overall survival and time to first therapy as well as more proliferation and less apoptosis. A quantification of the number of cells showing a genetic abnormality should, therefore, be included in the study of the prognostic factors of B-cell chronic lymphoid leukemia.

Incidence and Prognostic Impact of Secondary Cytogenetic Aberrations in a Series of 145 Patients with Mantle Cell Lymphoma

Mantle cell lymphoma (MCL) is a mature B‐cell neoplasm with an aggressive behavior, characterized by the t(11;14)(q13;q32). Several secondary genetic abnormalities with a potential role in the oncogenic process have been described. Studies of large MCL series using conventional cytogenetics, and correlating with proliferation and survival, are scarce. We selected 145 MCL cases at diagnosis, displaying an aberrant karyotype, from centers belonging to the Spanish Cooperative Group for Hematological Cytogenetics. Histological subtype, proliferative index and survival data were ascertained. Combined cytogenetic and molecular analyses detected CCND1 translocations in all cases, mostly t(11;14)(q13;q32). Secondary aberrations were present in 58% of patients, the most frequent being deletions of 1p, 13q and 17p, 10p alterations and 3q gains. The most recurrent breakpoints were identified at 1p31‐32, 1p21‐22, 17p13, and 1p36. Aggressive blastoid/pleomorphic variants displayed a higher karyotypic complexity, a higher frequency of 1p and 17p deletions and 10p alterations, a higher proliferation index and poor survival. Gains of 3q and 13q and 17p13 losses were associated with reduced survival times. Interestingly, gains of 3q and 17p losses added prognostic significance to the morphology in a multivariate analysis. Our findings confirm previous observations indicating that proliferation index, morphology and several secondary genetic alterations (3q gains and 13q and 17p losses) have prognostic value in patients with MCL. Additionally, we observed that 3q gains and 17p losses detected by conventional cytogenetics are proliferation‐independent prognostic markers indicating poor outcome.

Trisomies 8 and 20 in desmoid tumors

A nonrandom occurrence of trisomy 8 and of trisomy 20 in desmoid tumors has been recently reported. The finding of trisomy 8 in nondividing desmoid tumor cells by in situ hybridization prompted us to evaluate, in a similar way, the occurrence of trisomy 20 and the possible occurrence of both trisomies together because their co-existence was cytogenetically observed in a few cases. Double fluorescence in situ hybridization (FISH) with centromeric probes for chromosomes 8 and 20 was performed on 16 single cell suspensions of desmoid tumors. FISH confirmed the occurrence of trisomy 8 or 20 in a single cell suspension of desmoid tumors. Both individual trisomies, and even more their association in the same cells, are rare to extremely rare in solid tumors in general and in mesenchymal tumors in particular, and are only known to occur in infantile fibrosarcoma.

Germinal centre protein HGAL promotes lymphoid hyperplasia and amyloidosis via BCR-mediated Syk activation

The human germinal centre associated lymphoma (HGAL) gene is specifically expressed in germinal centre B-lymphocytes and germinal centre-derived B-cell lymphomas, but its function is largely unknown. Here we demonstrate that HGAL directly binds Syk in B-cells, increases its kinase activity upon B-cell receptor stimulation and leads to enhanced activation of Syk downstream effectors. To further investigate these findings in vivo, HGAL transgenic mice were generated. Starting from 12 months of age these mice developed polyclonal B-cell lymphoid hyperplasia, hypergammaglobulinemia and systemic reactive AA amyloidosis, leading to shortened survival. The lymphoid hyperplasia in the HGAL transgenic mice are likely attributable to enhanced B-cell receptor signalling as shown by increased Syk phosphorylation, ex vivo B-cell proliferation and increased RhoA activation. Overall, our study shows for the first time that the germinal centre protein HGAL regulates B-cell receptor signalling in B-lymphocytes which, without appropriate control, may lead to B-cell lymphoproliferation.

Detection of the Mbcr/abl translocation in chronic myeloid leukemia by fluorescence in situ hybridization: comparison with conventional cytogenetics and implications for minimal residual disease detection.

The correlation between the detection of the Philadelphia chromosome by conventional cytogenetics and the identification of Mbcr/abl translocation by fluorescence in situ hybridization (FISH) in both metaphase and interphase cells is prospectively analyzed in a group of 21 chronic myeloid leukemia (CML) patients. To gain insight into the sensitivity and specificity of the detection of the bcr/abl translocation by FISH, a group of 10 healthy volunteers was also studied. Our results show that for the detection of bcr/abl translocation in CML patients, FISH is more sensitive than conventional cytogenetics because it detects significantly higher proportions of cells carrying the translocation both in metaphase (P < .0002) and interphase nuclei (P < .003). Moreover, in the metaphases of the controls analyzed, no bcr/abl+ chromosome was detected that makes the colocalization of bcr and abl signals in the CML patients highly specific. Conversely, in control interphase nuclei, a small proportion of cells (ranging between 0% and 3%, mean value of 1.7% +/- 0.9%) displaying colocalization of both signals is usually detected. This limits, at least for the moment, the routine use of FISH for the detection of minimal residual disease in CML patients at levels lower than 10(-1).

Molecular Characterization of Chronic Lymphocytic Leukemia Patients with a High Number of Losses in 13q14

Background Patients with chronic lymphocytic leukemia and 13q deletion as their only FISH abnormality could have a different outcome depending on the number of cells displaying this aberration. Thus, cases with a high number of 13q- cells (13q-H) had both shorter overall survival and time to first therapy. The goal of the study was to analyze the genetic profile of 13q-H patients. Design and Methods: A total of 102 samples were studied, 32 of which served as a validation cohort and five were healthy donors. Results Chronic lymphocytic leukemia patients with higher percentages of 13q- cells (>80%) showed a different level of gene expression as compared to patients with lower percentages (<80%, 13q-L). This deregulation affected genes involved in apoptosis and proliferation (BCR and NFkB signaling), leading to increased proliferation and decreased apoptosis in 13q-H patients. Deregulation of several microRNAs, such as miR-15a, miR-155, miR-29a and miR-223, was also observed in these patients. In addition, our study also suggests that the gene expression pattern of 13q-H cases could be similar to the patients with 11q- or 17p-. Conclusions This study provides new evidence regarding the heterogeneity of 13q deletion in chronic lymphocytic leukemia patients, showing that apoptosis, proliferation as well as miRNA regulation are involved in cases with higher percentages of 13q- cells.